Ammonium sulphate

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Description of key information

There are no studies available on the effects of ammonium sulfate on fertility and development.

Based on data from a similar ammonium compound (diammonium phosphate), which has been tested up to 1500 mg/kg bw in a screening study according to OECD TG 422 in rats it can be concluded that ammonium ions up to the dose tested have no negative effects on fertility (NOAEL=1500 mg/kg bw/day).
In the 13-week feeding study of ammonium sulfate with rats, no histological changes of testes were observed up to 1792 mg/kg bw/day. The ovaries were not examined (NOAEL= 1792 mg/kg bw/day). Fully valid studies with sulfate on fertility are not available.
In a combined chronic oral / carcinogenicity study all gonads of the males and females were examined. No substance-related histopathological changes were observed up to 3% in the feed (NOAEL=1288.2 mg/kg bw/day for males, and 1371.4 mg/kg bw/day for females)
In a limited study (pretreatment time short, low number of animals, no fertility indices measured) where female mice were treated with up to ca. 6550 mg sulfate/kg bw (as sodium sulfate), no effects on litter size were found.
Data on purity of the respective test material are lacking in all of the studies. However, since no adverse reproductive effects or histopathological changes of the gonads were observed, this is considered not to significantly impair the evaluation of the reproductive toxicity of the test substance.
The overall conclusion is that ammonium sulfate does not present a hazard to fertility.

Link to relevant study records

Reference

Reference 1

Endpoint:

screening for reproductive / developmental toxicity

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

2001-12-28 to 2002-10-25

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Please refer to attached "Read-across justification" in IUCLID section 13.
In accordance to Regulation (EC) No 1907/2006 Annex XI 1.5. testing does not appear scientifically necessary if a read-across approach can be performed to common breakdown products. Ammonium sulfate dissociates into NH4+ and sulfate ions. Sulfate is a normal body and nutritional component and is regulated within the body. NH4+ ions are immediately transformed into urea by the liver and do not exist in the blood in relevant amounts unless in case of liver failure.

Reason / purpose for cross-reference:

read-across: supporting information

Qualifier:

according to guideline

Guideline:

OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)

Version / remarks:

adopted 1996-03-22

Deviations:

no

GLP compliance:

yes

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
male and CD rats, Crl:CD (TM) (SD)IGS BR strain
- Source: Charles River (UK) Limited, Margate, Kent England
- Age at study initiation: 7 - 8 weeks
- Weight at study initiation: Males: 298 - 386 g; Females: 191 - 263 g
- Housing: singly in RB3 modified cages
- Diet (e.g. ad libitum): Rat and Mouse No. 1 Maintenance Diet (Special Diets Services Ltd., Witham, Essex, England)
- Water (e.g. ad libitum): tap water from public supply
- Acclimation period: 16 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 23 °C
- Humidity (%): 40 - 70 %
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 2001-12-12 To: 2002-02-10 (necropsy)

Route of administration:

oral: gavage

Vehicle:

water

Details on exposure:

Dosing volume: 10 ml/kg bw for each group

Details on mating procedure:

All 10 males in each group (toxicity and reproductive subgroups) were mated with the 10 reproductive subgroup females after all animals had received 2 weeks of treatment. Pairing was on a one-to-one basis within treatment groups for up to 2 weeks, although all animals mated and were separated within 1 week.
Each morning following pairing the trays beneath the cages were checked for ejected copulation plugs and a wet vaginal smear was prepared from each female and examined for the presence of spermatozoa. The day on which a sperm positive vaginal smear or at least three copulation plugs were found was designated Day 0 of gestation. Once mating had been confirmed, males and females were separated and the males were returned to their normal group housing.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Samples of each formulation prepared for administration during Weeks l, 4 and 6 of the study were analysed for test material content and found to be satisfactory.

Duration of treatment / exposure:

Exposure period: not clearly defined
Premating exposure period (males): 2 weeks
Premating exposure period (females): 2 weeks
Duration of test: not clearly defined
See freetext below

Frequency of treatment:

once daily

Dose / conc.:

250 mg/kg bw/day (actual dose received)

Dose / conc.:

750 mg/kg bw/day (actual dose received)

Dose / conc.:

1 500 mg/kg bw/day (actual dose received)

No. of animals per sex per dose:

5 males and 5 females per group (toxicity subgroups);
5 males and 10 females per group (reproductive subgroups)

Control animals:

yes, concurrent vehicle

Parental animals: Observations and examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: once weekly
After removal from the home cage, animals were assessed for physical condition and behaviour during handling and after being placed in a standard arena. During these examinations particular attention was paid to possible signs of neurotoxicity such as convulsions, tremor and abnormalities of gait or behaviour.

BODY WEIGHT: Yes
- Time schedule for examinations: Each male and toxicity subgroup female was weighed on the day that treatment commenced, weekly thereafter, and at necropsy. Reproductive subgroup females were weighed on the first day of treatment, weekly until pairing and on Days 0, 7, 14, 17 and 20 after mating, and Days 1 and 4 of lactation, and at necropsy.

FOOD CONSUMPTION:
Food consumption for the males and toxicity subgroup females was recorded weekly throughout the study with the exception of the males whilst in pairing with the reproductive subgroup females. Food consumption for the reproductive subgroup females was recorded weekly before pairing then on Days 0, 7, 14, 17 and 20 after mating and Days 1 and 4 of lactation.
No food consumption was recorded for toxicity subgroup males or reproductive subgroup animals during pairing.

WATER CONSUMPTION: No data

OTHER:

FUNCTIONAL OBSERVATION BATTERY
All toxicity subgroup animals were subjected to FOB evaluation on one occasion following 4 weeks of treatment. Animals were not necessarily all tested on the same day but the number of animals was balanced across the groups on each day of testing. FOB examinations included:
- Sensory reactivity and grip strength
- Motor activity

HAEMATOLOGY AND BLOOD CHEMISTRY
During Week 5, following the FOB examinations, blood samples were obtained from the orbital sinus of all animals in the toxicity subgroup, following overnight deprivation of food. Samples were collected under light general anaesthesia (using Isoflurane) following overnight deprivation of food.
Parameters examined:
Haematocrit, Haemoglobin, Red blood cell count, Mean cell haemoglobin, Mean cell haemoglobin concentration, Mean cell volume, Total white cell count, Differential WBC count (Neutrophils, Lymphocytes, Eosinophils, Basophils, Monocytes, Large unstained cells, Platelet count), Reticulocyte count, Blood film, Prothrombin time, Activated partial thromoblastin time
Alkaline phosphatase, Alanine aminotransferase, Aspartate aminotransferase, Gamma-glutamyl transpeptidase, Total Bilirubin, Urea, Creatinine, Glucose, Total cholesterol, Sodium, Potassium, Chloride, Calcium, Inorganic Phosphorus, Magnesium, Total protein, Albumin

Litter observations:

All offspring were examined at approximately 24 hours after birth (Day 1) and the number of offspring born (live and dead) was recorded. Live offspring were individually identified within each litter by toe tattoo and any clinical observations were recorded.
Litters were observed daily for evidence of abnormal appearance or behaviour. Daily records were maintained of mortality and consequent changes in litter size. Where practical, any offspring found dead were subjected to a macroscopic examination as soon as possible. The offspring were sexed at individual weighing on Day I and Day 4 of age.

Postmortem examinations (parental animals):

GROSS PATHOLOGY
Toxicity subgroup animals were sacrificed during week 6 of treatment. The following tissues were retained, weighed and examined:
adrenals, aorta, brain, caecum, colon, duodenum, epididymis, eyes, heart, ileum, jejunum, kidneys, liver, lungs including bronchi, lymph nodes (mandibular, mesenteric, regional to masses), mammary area, oesophagus, optic nerves, ovaries, pancreas, pituitary, prostrate, rectum, salivary glands, sciatic nerves, seminal vesicles, skin, spinal cord, spleen, sternum with bone marrow, stomach, testes, thymus, thyroid with parathyroid, trachea, urinary bladder, uterus with cervix, vagina.

Reproductive subgroup males were killed after the toxicity subgroup animals. The testes, epididymides, seminal vesicles with coagulating gland and prostate were retained for each animal. Reproductive subgroup females were killed on Day 4 of lactation. The number of implantation sites was recorded and the ovaries, uterus with oviducts and cervix, vagina, pituitary and mammary tissue retained. Abnormal tissues were also retained.

HISTOPATHOLOGY
The tissues mentioned above from control and high dose group animals of the toxicity subgroups were examined histopathologically. Histology for reproductive subgroup animals was restricted to the retained reproductive organs and abnormalities observed at macroscopic necropsy.

Postmortem examinations (offspring):

Offspring of the reproductive sub group females were killed on Day 4 of age and subjected to a macroscopic necropsy.

CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS)
Treatment at all dosages was well tolerated and there were no treatment-related deaths.
A dosage dependent increase in transient post-dosing salivation was apparent, which was considered to be due to the palatability of the test formulations rather than toxicity. A dosage-dependent increase in the number of animals with reddening of the extremities was also apparent mainly during the early stages of treatment.

BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS)
Body weight gain and food consumption of males at 1500 mg/kg bw/day appeared to be suppressed when compared with the control group, such that gain between weeks 0-5 for this group was 78% of controls. The body weight gain for reproductive subgroup females receiving 1500 mg/kg bw/day was reduced during the first week of gestation, after which the values returned to levels comparable with the control.

REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS)
Mating performance and fertility were unaffected by treatment, and parental treatment had no apparent effect on the offspring to day 4 of age.

ORGAN WEIGHTS (PARENTAL ANIMALS)
Relative kidney and liver weights for females at 1500 mg/kg bw/day were greater than in the control group, but there were no histological changes associated.

GROSS PATHOLOGY, HISTOPATHOLOGY (PARENTAL ANIMALS)
A number of treated animals at 750 and 1500 mg/kg bw/day exhibited horizontal banding on the incisors at necropsy; histological processing of these tissues failed to detect any change in the areas examined suggesting that the banding was restricted to the enamel of the teeth. The only histological findings related to treatment were the inflammatory/degenerative stomach changes in all treated groups that were considered likely to have arisen due to an irritant effect of the test formulations.

OTHER FINDINGS (PARENTAL ANIMALS)

HAEMATOLOGY, CLINICAL CHEMISTRY
Some treatment-related effects on hematology were evident (reduction in activated partial thromboplastin time for males at 750 and 1500 mg/kg bw/day, a non dosage-dependent elevation of alkaline phosphatase levels at 750 and 1500 mg/kg bw/day, reduced glucose and phosphorous levels at 1500 mg/kg bw/day, a dosage-dependent reduction in total protein at 750 and 1500 mg/kg bw/day with a slight elevated albumin/globulin ratio at the top dosage. Changes in females were limited to a decrease in phosphorous levels and a non-significant increase in alkaline phosphatase level at
1500 mg/kg bw).

BEHAVIOUR
There were no changes apparent at behavioral testing.

Key result

Dose descriptor:

NOAEL

Remarks:

systemic toxicity

Sex:

male/female

Basis for effect level:

other: Based on the inflammatory/degenerative stomach changes recorded at histopathological examination and extending to the lowest dosage.

Remarks on result:

not determinable

Remarks:

no NOAEL identified

Parental treatment had no apparent effect on the offspring to day 4 of age.

Key result

Dose descriptor:

NOAEL

Remarks:

developmental toxicity

Generation:

F1

Effect level:

1 500 mg/kg bw/day (actual dose received)

Sex:

male/female

Basis for effect level:

other: Highest dose level applied.

Key result

Dose descriptor:

NOAEL

Remarks:

reproductive toxicity

Generation:

F1

Effect level:

1 500 mg/kg bw/day (actual dose received)

Sex:

male/female

Basis for effect level:

other: Highest dose level applied.

Reproductive effects observed:

not specified

Tabular Summary of effects on reproduction /development

Observations Dosage (mg/kg/day)	0 (ctr)	250	750	1500
Pairs started (N)	10	10	10	10
Females showing evidence of copulation (N)	10	10	10	10
Females achieving pregnancy (N)	9	10	10	10
Conceiving days 1 -5 (N)	9	10	10	10
Conceiving days 6 -14 (N)	0	0	0	0
Pregnancy=21.5 days (N)	0	0	0	1
Pregnancy=22 days (N)	6	9	8	4
Pregnancy=22.5 days (N)	3	0	2	4
Pregnancy=23 days (N)	0	1	0	1
Dams with live young born (N)	9	10	10	10
Dams with live young at day 4 pp (N)	9	10	10	10
Implants/dam (mean)	15.7	15.7	14.1	15.4
Live pubs/dam at birth (mean)	14.8	14.6	12.7	14.0
Live pubs/dam at days 4 (mean)	14.6	14.3	12.7	14.0
Sex ratio (%m) at birth (mean)	54.2	52.7	50.0	48.5
Sex ratio (m/f) at day 4 (mean)	54.2	53.0	50.0	48.5
Male pub weight at birth (mean)	6.4	6.3	6.6	6.3
Male pub weight at day 4 (mean)	8.7	8.5	9.2	8.7
Female pup weight at birth (mean)	5.9	6.0	6.1	6.0
Female pup weight at day 4 (mean)	8.2	7.9	8.6	8.4
Abnormal Pups: Necropsy finding unremarkable
Loss of offspring
Post implantation survival index	95.2	93.5	90.0	94.8
Live birth index	99.3	99.4	100.0	95.9
Viability index	98.6	98.2	100.0	100.0

Reference 2

Endpoint:

one-generation reproductive toxicity

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

no data

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Justification for type of information:

Please refer to attached "Read-across justification" in IUCLID section 13.
In accordance to Regulation (EC) No 1907/2006 Annex XI 1.5. testing does not appear scientifically necessary if a read-across approach can be performed to common breakdown products. Ammonium sulfate dissociates into NH4+ and sulfate ions. Sulfate is a normal body and nutritional component and is regulated within the body. NH4+ ions are immediately transformed into urea by the liver and do not exist in the blood in relevant amounts unless in case of liver failure.

Reason / purpose for cross-reference:

read-across: supporting information

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 415 [One-Generation Reproduction Toxicity Study (before 9 October 2017)]

Deviations:

yes

Remarks:

low numbers of animals per dose group, only females were treated, no fertility indices were measured.

Principles of method if other than guideline:

10 females were treated with sodium sulfate, the males were untreated.
Sixty random-bred ICR female virgin mice were used. Mice were assigned randomly to one of six water treatments (control, Na control, 4 dose levels of sodium sulfate). All groups contained equal levels of Na, which was maintained by varying the amount of sodium bicarbonate added. The treated water was available ad libitum beginning 1 wk prior to breeding and was continued throughout the experiment. After 1 wk of acclimation to the treatments, a male mouse that had received tap water was paired randomly with each female mouse. The females were checked every 24 hours in the morning for  the presence of a vaginal plug. After a vaginal plug was observed, the  male mouse was removed and the female was weighed. Water consumption was measured daily during the 2nd and 3rd wk of gestation, and the 1st and 2nd wk of lactation. At parturition, the dams were weighed and litter size was recorded. The litters then were standardized to eight pups per litter. At 21 days postpartum, the pups were weaned and the litters and dams were weighed individually. The dams were then rebred at first estrus immediately following weaning. This procedure was carried out over two parities. Only animals that whelped during each parity were used in the analysis. Thus, the number of dams per dose group in the first parity  was 4-9, in the second parity it was only 4. Fertility indices were not  measured.

GLP compliance:

not specified

Limit test:

no

Species:

mouse

Strain:

ICR

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
random-bred virgin female ICR mice
- Housing: individually in stainless steel cages
- Diet (ad libitum): commercially prepared breeder block
- Water (ad libitum): dosing solution or control water
- Acclimation period: 1 week

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 23.6 °C
- Humidity (%): 55%
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: drinking water

Vehicle:

water

Details on mating procedure:

cohabitation with untreated males
- M/F ratio per cage: 1/1
- Length of cohabitation: 24 h
- Proof of pregnancy: vaginal plug

Duration of treatment / exposure:

Exposure period: from one week prior to breeding until study end (day 21 of second parity)
Premating exposure period (males): no treatment (tap water ad lib)
Premating exposure period (females): 1 week

Frequency of treatment:

continuously in the drinking water

Dose / conc.:

625 mg/L drinking water

Dose / conc.:

1 250 mg/L drinking water

Dose / conc.:

2 500 mg/L drinking water

Dose / conc.:

5 000 mg/L drinking water

No. of animals per sex per dose:

10 females per group

Control animals:

yes, concurrent vehicle

Statistics:

Least squares mean analysis of variance (SAS, 1982).
Student's t-test was used to determine the difference in water consumption.

Key result

Dose descriptor:

NOAEL

Effect level:

5 000 mg/L drinking water

Sex:

female

Basis for effect level:

mortality

Key result

Dose descriptor:

NOAEL

Effect level:

5 000 mg/L drinking water

Sex:

male/female

Basis for effect level:

other: no effects observed

Remarks on result:

not determinable

Key result

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

5 000 mg/L drinking water

Sex:

male/female

Basis for effect level:

other: no effects observed

Reproductive effects observed:

not specified

The effect of sulfate treatment on water consumption (first parity)

Sulfate level ppm	No. of mice	Period
		7 to 14 d gestation	14 to 21 d gestation	0 to 7 d lactation	7 to 14 d lactation
		ml/wk
0 (ddH2O)	6	70.3*	95.3*	129.8*	233.8*
0^§	6	113.5#	160.3#	254.5#	369.0#
625	4	82.5#	108.0#	159.5#	271.0#
1250	5	81.0#	106.0#	183.0#	284.5#
2500	9	107#	158.3#	217.8#	329.3#
5000	7	75.0#	98.0#	165.8#	270.0#
pooled means		88.2^f	120.9^g	185.1^h	292.9^i
± SE		± 8.0	± 9.4	± 19.6	± 21.8

*/# Sulfate treatment means with same colum with different superscript letters differ from the O sulfate, Na control treatment (p<0.05)

^§ O sulfate, na control treatment

^f,g Means between gestational weeks differ (p<0.01)

^h,i Means between lactational weeks differ (p<0.01)

The effect of sulfate treatment on water consumption (second parity)

Sulfate level ppm	No. of mice	Period
		7 to 14 d gestation	14 to 21 d gestation	0 to 7 d lactation	7 to 14 d lactation
		ml/wk
0 (ddH2O)	4	86.8	101.3*	146.8*	237.5*
0^§	4	120.8	159.5#	282.3#	411.3#
625	4	90.8	125.8#	163.3#	302.3#
1250	4	99.8	165.5#	257.0#	377.5#
2500	4	125.8	168.0#	247.5#	411.3#
5000	4	85.8	117.0#	175.8#	303.3#
pooled means		101.6^f	139.5^g	212.1^h	340.5^i
± SE		± 10.4	± 16.1	± 23.9	± 35.7

*/# Sulfate treatment means with same colum with different superscript letters differ from the O sulfate, Na control treatment (p<0.05)

^§O sulfate, Na control treatment

^f,gMeans between gestational weeks differ (p<0.01)

^h,iMeans between lactational weeks differ (p<0.01)

The effects of sulfate treatment on gestational(a) and lactational(b) weight gain of dams (both parities)

Sulfate level ppm	Weight gain^cin g
	gestation	lactation	gestation	lactation
	first parity		second parity
0 (ddH2O)	2.5	5.0	1.1	2.2
0^§	3.1	4.9	1.5	2.7
625	1.7	3.5	0.9	1.7
1250	3.7	7.5	1.3	2.5
2500	3.7	7.4	2.1	3.9
5000	3.1	6.2	1.6	3.1
SE	0.5	1.1	0.4	0.7

^aGestational gain = (weight after parturition-weight at conception)

^bLactational gain =(weight at 21d postpartum-weight after parturition)

^c Sulfate treatment means within columns do not differ from the O sulfate, sodium control treatment (p>0.01)

^§O sulfate, Na control treatment

The effects of sulfate treatment on litter size (a) at weaning and litter weaning weight (both parities)

Sulfate level ppm
	pups/litter	weaning ^c, wt, g	pubs/litter	weaning ^c wt,g
	first parity^b		second partity^b
0 (ddH2O)	8.0	123.3	11.3	129.7
0^§	8.3	128.1	13.0	130.6
625	8.8	114.3	13.3	126.9
1250	7.8	130.0	13.5	141.2
2500	8.0	132.7	13.0	144.2
5000	8.5	125.9	15.5	130.4
SE	0.6	6.2	1.3	3.8^f

^a total number born and number born alive were identical because no mortality was detected at parturition

^b sulfate treatment means within columns do not differ from the O sulfate,Na control treatment (p>0.01).

^c weaning weights of the standardized /(eight pups) litters

^§O sulfate, Na control treatment

^f Quadratic response within sulfate treatment (p< 0.005)

Reference 3

Endpoint:

reproductive toxicity, other

Remarks:

Cancer study 104 weeks

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

no data

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: cancer study, histopathological results of gonades

Principles of method if other than guideline:

These investigations were part of a 104-week oral long term toxicity study.

GLP compliance:

not specified

Limit test:

no

Species:

rat

Strain:

Fischer 344/DuCrj

Sex:

male/female

Details on test animals or test system and environmental conditions:

see chapter 7.5.1 and 7.7

Route of administration:

oral: feed

Vehicle:

unchanged (no vehicle)

Details on exposure:

see chapter 7.5.1 and 7.7

Duration of treatment / exposure:

104 weeks

Frequency of treatment:

daily in feed

Dose / conc.:

564.2 mg/kg bw/day (actual dose received)

Remarks:

males (1.5% nominal in diet)

Dose / conc.:

1 288 mg/kg bw/day (actual dose received)

Remarks:

males (3% nominal in diet)

Dose / conc.:

649.9 mg/kg bw/day (actual dose received)

Remarks:

females (1.5% nominal in diet)

Dose / conc.:

1 371.4 mg/kg bw/day (actual dose received)

Remarks:

females (3% nominal in diet)

No. of animals per sex per dose:

control: 50 males and 48 females
1.5 %: 48 males and 48 females
3.0%. 50 males and 50 females

Control animals:

yes, concurrent no treatment

Parental animals: Observations and examinations:

OTHER: see chapter 7.5.1 and 7.7

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

see "Details on results"

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

no effects observed

CLINICAL SIGNS AND MORTALITY
In the chronic toxicity study, no mortality was found in any groups throughout the treatment period.
In the carcinogenicity study, the survival rate of control, 1.5% and 3.0% groups were 88%, 78% and 76%, respectively, for males, and 76%, 80% and 80%, respectively, for females, and no significant differences were observed among the groups.

BODY WEIGHT AND WEIGHT GAIN
No test substance-related change in the body weights was found.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study)
A tendency for increase of food intake in the male 3.0% group in the chronic toxicity study was noted.

HAEMATOLOGY
No significant variation was found.

CLINICAL CHEMISTRY
No dose related alteration was found.

ORGAN WEIGHTS
Absolute and relative kidney weights were increased or showed a tendency for increase at 3.0% in both sexes in the chronic toxicity study. Absolute spleen weights were decreased and relative liver weights were increased in the 3.0% male dose group. No dose-related changes were found in the other organs.

GROSS PATHOLOGY
There were no obvious macroscopic findings in any group in either the chronic toxicity or carcinogenicity studies, except for massive, nodular or focal lesions suggesting neoplastic change in the carcinogenicity study.

HISTOPATHOLOGY:
In the carcinogenicity study, non-neoplastic and neoplastic lesions were noted in the control and treatment groups, with no significant inter-group difference in their incidences or severity.

Key result

Dose descriptor:

NOAEL

Effect level:

1 288.2 mg/kg bw/day (actual dose received)

Sex:

male

Basis for effect level:

other: histopathology of gonades

Key result

Dose descriptor:

NOAEL

Effect level:

1 371.4 mg/kg bw/day

Sex:

female

Basis for effect level:

other: histopathology of gonades

Remarks on result:

not measured/tested

Reproductive effects observed:

not specified

There were no significant changes in absolute and relative testes weight. Histopathological examination after 104 weeks dietary treatment revealed no significant changes of the testes, ovaries, uterus, vagina or mammary gland up to the highest dose (NOAEL reprotoxicity: females: 1371.4 mg/kg and males 1288.2 mg/kg).

Reference 4

Endpoint:

reproductive toxicity, other

Remarks:

subchronic oral toxicity study

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

no data

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Ovaries not weighted and examined.

Principles of method if other than guideline:

no data
These investigations were part of a 13-week oral toxicity study.

GLP compliance:

not specified

Limit test:

no

Species:

rat

Strain:

Fischer 344/DuCrj

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
male and female F344/DuCrj (SPF) rats
- Source: Charles River Japan
- Age at study initiation: 5 weeks
- Housing: 5 per cage
- Diet (ad libitum): basal diet (CRF-1) without or with the test substance added
- Acclimation period: 1 week

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 23-25°C
- Humidity (%): 50-60%
- Air changes (per hr): 18
- Photoperiod (hrs dark / hrs light): 12/12

No further data

Route of administration:

oral: feed

Vehicle:

unchanged (no vehicle)

Details on exposure:

DIET PREPARATION
- Storage temperature of food: room temperature
No further data

Details on mating procedure:

no mating

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

13 weeks

Frequency of treatment:

continuously in the diet

Dose / conc.:

220 mg/kg bw/day (actual dose received)

Remarks:

males (0.38% nominal in diet)

Dose / conc.:

441 mg/kg bw/day (actual dose received)

Remarks:

males (0.75% nominal in diet)

Dose / conc.:

886 mg/kg bw/day (actual dose received)

Remarks:

males (1.5% nominal in diet)

Dose / conc.:

1 792 mg/kg bw/day (actual dose received)

Remarks:

males (3% nominal in diet)

Dose / conc.:

239 mg/kg bw/day (actual dose received)

Remarks:

females (0.38% nominal in diet)

Dose / conc.:

484 mg/kg bw/day (actual dose received)

Remarks:

females (0.75% nominal in diet)

Dose / conc.:

961 mg/kg bw/day (actual dose received)

Remarks:

females (1.5% nominal in diet)

Dose / conc.:

1 975 mg/kg bw/day (actual dose received)

Remarks:

females (3% nominal in diet)

No. of animals per sex per dose:

10 males and 10 females per group

Control animals:

yes, plain diet

Details on study design:

- Dose selection rationale: The dose levels were set on the basis of results from a previous 2-week study with a dose level of 5% (no further details reported).
- Rationale for animal assignment: random

Parental animals: Observations and examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily
- Cage side observations checked: mortality, behaviour

DETAILED CLINICAL OBSERVATIONS: No data

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

FOOD CONSUMPTION: Yes
- Time schedule for examinations: weekly

FOOD EFFICIENCY: No

WATER CONSUMPTION: No

OTHER:

HAEMATOLOGY and CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: Blood was collected from the aorta abdominalis post mortem (during necropsy)
- How many animals: all animals
- Parameters examined:
--erythrocyte count, hemoglobin, hematocrit, mean cell volume, mean corpuscular hemoglobin content, mean corpuscular hemoglobin concentration, platelet count, leukocyte count
-- total protein, albumin/globulin ratio, albumin, total cholesterol, urea nitrogen, sodium, chloride, potassium, calcium, inorganic phosphorus, glutamat-oxaloacetic transaminase, glutamate-pyruvat transaminase, alkaline phosphatase

Postmortem examinations (parental animals):

GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes

Following section, the animals were exsanguinated. Brain, heart, lungs, liver, kidneys, adrenals, spleen, testes, and thymus were weighed, fixed (10% neutral buffered formalin), sectioned, stained (hematoxyline and eosin) and examined histologically.
Ovaries were not examined.

Statistics:

Bartlett' test (organ weights), Kruskal-Wallis test, and parametric Dunnett and Scheffe tests.

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

see "Details on results"

Mortality:

no mortality observed

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

see "Details on results"

Food consumption and compound intake (if feeding study):

no effects observed

Food efficiency:

not examined

Ophthalmological findings:

not examined

Haematological findings:

effects observed, treatment-related

Description (incidence and severity):

see "Details on results"

Urinalysis findings:

not examined

Behaviour (functional findings):

not examined

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

effects observed, non-treatment-related

CLINICAL SIGNS AND MORTALITY
No deaths were observed throughout the study.
Diarrhea was noted in males of the 3% group, starting one week after beginning of treatment and persisting until termination.

BODY WEIGHT AND WEIGHT GAIN
Body weight gain was significantly decreased in 3% males when compared with controls.
Final mean body weights were significantly decreased in males receiving 0.38%, 1.5% and 3%, and significantly increased in females of the same groups when compared with controls. The changes in the 0.38% group were not dose-related.


FOOD CONSUMPTION
There were no significant differences between the groups for both males and females.

HAEMATOLOGY and CLINICAL CHEMISTRY
The following, statistically significant and dose-related changes were observed:
- decreased leukocyte count (3% males)
- decreased erythrocyte count, hemoglobin, decreased hematocrit, and platelet count (3% females).
- increased mean corpuscular hemoglobin content and mean corpuscular hemoglobin concentration (3% females)
The same changes in hematological parameters were also observed in 1.5% males and females; however, these changes were not dose-related.
Blood serum analysis revealed some statistically significant alterations; however, again there was no dose-response.


ORGAN WEIGHTS
There were some statistically signficant changes in absolute and relative organ weights. However, these changes were not dose-related.

PATHOLOGY:
Histological examination revealed myofibrosis cordis (3% males), basophilia of the renal tubuli (3% males and females), and melanosis of the spleen (3% males and females). However, the incidence of these findings was not statistically significantly different from control.

According to the authors, with exception of the diarrhea observed in high dose males, none of these observation was considered to indicate obvious toxicity of the test substance.

Key result

Dose descriptor:

NOAEL

Effect level:

1 792 mg/kg bw/day (actual dose received)

Sex:

male

Basis for effect level:

other: histology of testis

Remarks on result:

not measured/tested

Reproductive effects observed:

not specified

There were no significant changes in absolute and relative testes weight. Histopathological examination revealed no significant changes of the testes. Ovaries were not examined.

Effect on fertility: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 500 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

OECD TG 422

Effect on fertility: via inhalation route

Endpoint conclusion:

no study available

Effect on fertility: via dermal route

Endpoint conclusion:

no study available

Additional information

There are no valid studies available in which ammonium sulfate has been tested for its effects on fertility and development. The following are available for the ammonium and the sulfate ion.

In a 13-week feeding study of ammonium sulfate with rats, no histological changes of testes were observed up to 1792 mg/kg bw/day. The ovaries were not examined (see. chapter also section 7.5.1; Takagi et al., 1999). In a combined chronic oral / carcinogenicity study all gonads of the males and females were examined. No substance-related histopathological changes were observed in the gonades up to 3% in feed (1288.2 mg/kg bw/day in males and 1371.4 mg/kg bw/day in females) (see chapter 7.5.1 and 7.7 Ota et al. 2006).

In a gavage study performed according to OECD TG 422 with diammonium phosphate (250, 750, 1500 mg/kg; purity of the test substance not stated) in CD rats (10 females and 5 males/group), no mortality was observed up to the highest tested dose level of 1500 mg/kg bw/day. Treatment of all dosages was well tolerated. A dosage dependant increase in transient post-dosing salivation was apparent, which was considered to be due to the palatability of the test formulations rather than toxicity. A dosage-dependant increase in the number of animals with reddening of the extremities was also apparent mainly during the early stages of treatment. The body weight gain and food consumption of males at 1500 mg/kg bw/day appeared to be suppressed when compared with Control, such that gain between Weeks 0 -5 for this group was 78% of Control.

Body weight gain for reproductive subgroup females receiving 1500 mg/kg bw/day was reduced during the first week of gestation, after which values returned to Control-comparable levels through to termination at Day 4 of lactation.

The findings of the haematology investigations were largely unremarkable; the only treatment-related change evident was a reduction in activated partial thromboplastin time for males at 750 or 1500 mg/kg bw/day. There was slightly greater variability in blood chemistry parameters, and the following were blood chemistry changes for males for which an effect of treatment could not be discounted: a non dosage-dependant elevation of alkaline phosphatase levels at 750 and 1500 mg/kg bw/day; reduced glucose and phosphorous levels at 1500 mg/kg bw/day; a dosage-dependant reduction in total protein at 750 and 1500 mg/kg bw/day with a slightly elevated albumin/globulin ratio at the top dosage. Changes in females were limited to a decrease in phosphorous levels and a non-significant increase in alkaline phosphatase levels at 1500 mg/kg bw/day.

There were no changes apparent at behavioural testing that were consistent with a clear effect of treatment.

Mating performance and fertility were unaffected by treatment and parental treatment had no apparent effect on the offspring to Day 4 of age. Body weight relative kidney and liver weights for females at 1500 mg/kg/day were greater than Control, but these were not associated with histological changes.

A number of treated animals at 750 or 1500 mg/kg bw/day exhibited horizontal banding on the incisors at necropsy; histological processing of these tissues failed to detect any change in the areas examined suggesting that the banding was restricted to the enamel of the teeth. The only histological findings clearly related to treatment were the inflammatory/degenerative stomach changes in all treated groups that were considered likely to have arisen due to an irritant effect of the test formulations.

Bodyweight gain was temporarily reduced in the high dose group (in males week 0 - 5, in females week 1). No treatment-related signs of clinical toxicity were observed. Mating performance and fertility were unaffected by parental exposure to diammonium phosphate (The Weinberg Group, 2002).

Groups of 10 female ICR mice were given sodium sulfate in drinking water at levels of 0 (= sodium control), 625, 1250, 2500 or 5000 mg sulfate/l (ca. 250 - 850, 480 - 2040, 1270 - 4320, 1790 - 6560 mg sulfate/kg bw) beginning one week prior to breeding and up to 14 days during lactation. At day 21 p.p. the pups were weaned and the dams were rebred at first estrus immediately following weaning. Only animals that whelped during each parity were used in the analysis. The effective number of dams per group was low: 4 - 9 in the first parity, and 4 in the second parity. Control mice, receiving only distilled water, consumed significantly less water than mice receiving sulfate treatments, and sodium-control mice drank significantly more water than mice treated with sulfate. No differences were found in litter size, litter weaning weights, or gestational or lactational weight gain of the dams among sulfate treatments. No toxicity to the dams was found. Litters were not histopathologically examined. Fertility indices were not measured (Andres and Cline, 1989). The study is limited in that only females were treated, the number of dams per group was low and no fertility indices were measured. Moreover the pretreatment period was short.

Effects on developmental toxicity

Description of key information

Studies of developmental toxicity for ammonium sulfate are not available.

In the screening study according to OECD TG 422 with up to 1500 mg diammonium phosphate/kg bw, no effects on development have been detected in rats. Therefore, the NOAEL for developmental toxicity within the scope of these data was 1500 mg/kg/day

In another limited screening study with exposure of mice to a single dose of 2800 mg sodium sulfate/kg bw no macroscopic effects or adverse effects on body weight gain have been detected in the pups. In both studies fetuses were not examined histopathologically.

The overall conclusion is that there is no evidence that ammonium sulfate may present a risk of a developmentally toxic/teratogenic potential.

Link to relevant study records

Reference

Reference 1

Endpoint:

developmental toxicity

Remarks:

Combined Repeated Dose Toxicity Study with Reproduction/Developmental Toxicity Screening Test

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

2001-12-28 to 2001-10-25

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

other: OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Toxicity Screening Test)

Version / remarks:

adopted 1996-03-22

Deviations:

no

GLP compliance:

yes

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Details on test animals or test system and environmental conditions:

TEST ANIMALS
male and CD rats, Crl:CD (TM) (SD)IGS BR strain
- Source: Charles River (UK) Limited, Margate, Kent England
- Age at study initiation: 7 - 8 weeks
- Weight at study initiation: Males: 298 - 386 g; Females: 191 - 263 g
- Housing: singly in RB3 modified cages
- Diet (e.g. ad libitum): Rat and Mouse No. 1 Maintenance Diet (Special Diets Services Ltd., Witham, E
ssex, England)
- Water (e.g. ad libitum): tap water from public supply
- Acclimation period: 16 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 23 °C
- Humidity (%): 40 - 70 %
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12
IN-LIFE DATES: From: 2001-12-12 To: 2002-02-10 (necropsy)

Route of administration:

oral: gavage

Vehicle:

water

Details on exposure:

Dosing volume: 10 ml/kg bw for each group

Details on mating procedure:

All 10 males in each group (toxicity and reproductive subgroups) were mated with the 10 reproductive
subgroup females after all animals had received 2 weeks of treatment. Pairing was on a one-to-one basis
within treatment groups for up to 2 weeks, although all animals mated and were separated within 1 week
.
Each morning following pairing the trays beneath the cages were checked for ejected copulation plugs
and a wet vaginal smear was prepared from each female and examined for the presence of spermat
ozoa. The day on which a sperm positive vaginal smear or at least three copulation plugs were found was
designated Day 0 of gestation. Once mating had been confirmed, males and females were separated
and the males were returned to their normal group housing.

Duration of treatment / exposure:

Exposure period: not clearly defined
Premating exposure period (males): 2 weeks
Premating exposure period (females): 2 weeks
Duration of test: not clearly defined
See free text below

Frequency of treatment:

once daily

Dose / conc.:

250 mg/kg bw/day (actual dose received)

Dose / conc.:

750 mg/kg bw/day (actual dose received)

Dose / conc.:

1 500 mg/kg bw/day (actual dose received)

No. of animals per sex per dose:

5 males and 5 females per group (toxicity subgroups);
5 males and 10 females per group (reproductive subgroups)

Control animals:

yes, concurrent vehicle

Details on results:

CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS)
Treatment at all dosages was well tolerated and there were no treatment-related deaths.
A dosage dependent increase in transient post-dosing salivation was apparent, which was considered to
be due to the palatability of the test formulations rather than toxicity. A dosage-dependent increase in the
number of animals with reddening of the extremities was also apparent mainly during the early stages of
treatment.
BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS)
Body weight gain and food consumption of males at 1500 mg/kg bw/day appeared to be suppressed whe
n compared with the control group, such that gain between weeks 0-5 for this group was 78% of contr
ols. The body weight gain for reproductive subgroup females receiving 1500 mg/kg bw/day was reduced
during the first week of gestation, after which the values returned to levels comparable with the control.
REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS)
Mating performance and fertility were unaffected by treatment, and parental treatment had no apparent
effect on the offspring to day 4 of age.
ORGAN WEIGHTS (PARENTAL ANIMALS)
Relative kidney and liver weights for females at 1500 mg/kg bw/day were greater than in the control
group, but there were no histological changes associated.
GROSS PATHOLOGY, HISTOPATHOLOGY (PARENTAL ANIMALS)
A number of treated animals at 750 and 1500 mg/kg bw/day exhibited horizontal banding on the incisors
at necropsy; histological processing of these tissues failed to detect any change in the areas examined
suggesting that the banding was restricted to the enamel of the teeth. The only histological findings
related to treatment were the inflammatory/degenerative stomach changes in all treated groups that were
considered likely to have arisen due to an irritant effect of the test formulations.
OTHER FINDINGS (PARENTAL ANIMALS)
HAEMATOLOGY, CLINICAL CHEMISTRY
Some treatment-related effects on hematology were evident (reduction in activated partial thromboplasti
n time for males at 750 and 1500 mg/kg bw/day, a non dosage-dependent elevation of alkaline phospha
tase levels at 750 and 1500 mg/kg bw/day, reduced glucose and phosphorous levels at 1500 mg/kg bw/
day, a dosage-dependent reduction in total protein at 750 and 1500 mg/kg bw/day with a slight elevated
albumin/globulin ratio at the top dosage. Changes in females were limited to a decrease in phosphorous
levels and a non-significant increase in alkaline phosphatase level at 1500 mg/kg bw/day).
BEHAVIOUR
There were no changes apparent at behavioral testing.

Remarks on result:

other: based on inflammatory/degenerative stomach changes recorded at histopathological examination and extending to the lowest dosage investigated,no NOAEL for general toxicity could be established

Key result

Dose descriptor:

NOAEL

Effect level:

1 500 mg/kg bw/day (actual dose received)

Sex:

male

Basis for effect level:

other: Highest dose applied

Key result

Dose descriptor:

NOAEL

Effect level:

1 500 mg/kg bw/day (actual dose received)

Sex:

female

Basis for effect level:

other: Highest dose applied

Developmental effects observed:

no

Tabular Summary of effects on reproduction /development

Observations Dosage (mg/kg/day)	0 (ctr)	250	750	1500
Pairs started (N)	10	10	10	10
Females showing evidence of copulation (N)	10	10	10	10
Females achieving pregnancy (N)	9	10	10	10
Conceiving days 1 -5 (N)	9	10	10	10
Conceiving days 6 -14 (N)	0	0	0	0
Pregnancy=21.5 days (N)	0	0	0	1
Pregnancy=22 days (N)	6	9	8	4
Pregnancy=22.5 days (N)	3	0	2	4
Pregnancy=23 days (N)	0	1	0	1
Dams with live young born (N)	9	10	10	10
Dams with live young at day 4 pp (N)	9	10	10	10
Implants/dam (mean)	15.7	15.7	14.1	15.4
Live pubs/dam at birth (mean)	14.8	14.6	12.7	14.0
Live pubs/dam at days 4 (mean)	14.6	14.3	12.7	14.0
Sex ratio (%m) at birth (mean)	54.2	52.7	50.0	48.5
Sex ratio (m/f) at day 4 (mean)	54.2	53.0	50.0	48.5
Male pub weight at birth (mean)	6.4	6.3	6.6	6.3
Male pub weight at day 4 (mean)	8.7	8.5	9.2	8.7
Female pup weight at birth (mean)	5.9	6.0	6.1	6.0
Female pup weight at day 4 (mean)	8.2	7.9	8.6	8.4
Abnormal Pups: Necropsy finding unremarkable
Loss of offspring
Post implantation survival index	95.2	93.5	90.0	94.8
Live birth index	99.3	99.4	100.0	95.9
Viability index	98.6	98.2	100.0	100.0

Reference 2

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

no data

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Limited documentation, short exposure period, fetuses were not examined histopathologically

Qualifier:

no guideline followed

Principles of method if other than guideline:

Screening test with 55 chemicals

GLP compliance:

not specified

Species:

mouse

Strain:

ICR

Details on test animals or test system and environmental conditions:

TEST ANIMALS
adult barrier-reared ICR/SIM mice
- Source: Simonsen Laboratories (Gilroy, CA)
- Weight at study initiation: 32 - 36 g
- Housing: individually in 32 X 23 X 15 cm suspended polycarbonate shoebox cages
- Diet (ad libitum): Simonsen Custom Lab Diet 7
- Water (ad libitum): UV purified drinking water
Mice were either received from the supplier timed-pregnant or were bred in-house.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 - 24°C
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: gavage

Vehicle:

water

Duration of treatment / exposure:

days 8 through 12 of gestation

Frequency of treatment:

daily

Duration of test:

until day 3 post partum

Dose / conc.:

2 800 mg/kg bw/day (actual dose received)

No. of animals per sex per dose:

28 females per group

Control animals:

yes, concurrent vehicle

Statistics:

All analyses compared individual treatment groups and their concurrent controls. Maternal body weight gain during treatment (day 13 minus day 7 weight) was assessed by a two-tailed analysis of variance. A one-tailed analysis of variance was used to assess live and dead litter size data. The calculation of the average live litter size on days 1 and 3 and the average number of neonates found dead per litter on day 1 included only those litters that were born. Litters that were alive in utero at necropsy and those that were completely resorbed will be discussed as appropriate, but were not included in this average. A one-tailed Fisher's exact probability test was used to assess neonatal survival ratios. Neonatal body weight was assessed by a two-tailed analysis of variance. To correct for differences in litter size, the number of live neonates born was used as a covariant in the body weight analyses.

Key result

Dose descriptor:

NOAEL

Effect level:

> 2 800 mg/kg bw/day (actual dose received)

Basis for effect level:

other: maternal toxicity

Key result

Dose descriptor:

NOAEL

Effect level:

> 2 800 mg/kg bw/day (actual dose received)

Basis for effect level:

other: other:

Key result

Dose descriptor:

NOAEL

Effect level:

> 2 800 mg/kg bw/day (actual dose received)

Basis for effect level:

other: teratogenicity

Abnormalities:

not specified

Developmental effects observed:

not specified

No evidence of maternal toxicity (mortality, body weight gain), or increased resorption rates was found. The test substance had no effects on pup survival, and no adverse developmental effects were observed. When compared to vehicle controls, birth weight was significantly increased. All of the confirmed teratogens tested in this study resulted in decreased live-born litter size.

Table: Maternal and perinatal effects of sodium sulfate

Treatment

Maternal response

Perinatal response

Average neonatal weight (g +/- SD)

Compound

Dose (mg/kg/d)

No. dead/ No. treated

Average weight gain (g+/-SD)

No. of litters

Average no. of neonated/litter (+/- SD)

% survival days 1-3

day 1

day 3

2-day gain

born

resorbed

live day 1

dead day 1

live day 3

Vehicle

---

0/28

8.5 +/- 1.7

25

0

13.3 +/- 3.8

0.16 +/- 0.5

13.2 +/- 3.8

99

1.72 +/- 0.13

2.42 +/- 0.25

0.70 +/- 0.15

Test substance

2800

0/28

8.6 +/- 1.7

24

0

12.9 +/- 2.5

0.04 +/- 0.2

12.8 +/- 2.4

100

1.80 +/- 0.14 *

2.58 +/- 0.29

0.78 +/- 0.18

* p<0.05

Effect on developmental toxicity: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 500 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

OECD TG 422

Effect on developmental toxicity: via inhalation route

Endpoint conclusion:

no study available

Effect on developmental toxicity: via dermal route

Endpoint conclusion:

no study available

Additional information

No data are available on the developmental toxicity of ammonium sulfate.

In a gavage screening study performed according to OECD TG 422 with diammonium phosphate

(250, 750, 1500 mg/kg bw in rats, animals were paired 2 weeks after start of treatment), no mortality was observed. Some treatment-related effects on hematology parameters (at 750 and 1500 mg/kg bw) were evident, and bodyweight gain was temporarily reduced in the high dose group. No treatment-related signs of clinical toxicity were observed. Banding of the enamel of the incisors of the parent animals from 750 mg/kg bw. was reported, likely due to the inhibiting effect of phosphate on mineralisation of the teeth. Offspring was unaffected by parental exposure to diammonium phosphate. Viability up to day 4 pp. and macroscopic necropsy showed no effect on the pups. The NOAEL for reproductive/ developmental toxicity within the scope of these data was 1500 mg/kg/day (The Weinberg Group, 2002).

In a screening study, aqueous sodium sulfate was given by gavage to 28 time-pregnant ICR mice at a dose of 2800 mg/kg bw/day from gestation days 8 through 12. The dose was selected based on previous range-finding study in non-pregnant mice, 10 % mortality was expected. Mice were allowed to deliver, and neonates were macroscopically examined, counted, and weighed on day 1 and 3. No evidence of maternal toxicity or increased resorption rate was found. The chemical had no influence on pup survival, and no adverse developmental effects on external examination were observed. When compared to controls, birth weight was significantly increased (Seidenberg et al., 1986). Fetuses were not examined histologically, and the exposure period was short compared to a valid guideline study, therefore the study is of limited reliability.The NOAEL was determined to be >2800 mg/kg bw/day.

Justification for classification or non-classification

Classification, Labelling, and Packaging Regulation (EC) No 1272/2008

The available experimental test data are reliable and suitable for classification purposes under Regulation (EC) No 1272/2008. As a result the substance is not considered to be classified under Regulation (EU) No 1272/2008, as amended for the ninth time in Regulation (EU) No 2016/1179.